1. Antibodies
  2. Homo sapiens
  3. SARNP

ENCAB353OXX

Antibody against Homo sapiens SARNP

Homo sapiens
HepG2, K562
characterized to standards
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-209A
Lot ID
1
Characterized targets
SARNP (Homo sapiens)
Host
rabbit
External resources

Characterizations

SARNP (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
IP-WB analysis of K562 whole cell lysate using CIP29 specific antibody. Lane 1 is 2.5% of 0.5mg input lysate, lane 2 is 2.5% of supernatant after immunoprecipitation and Lane 3 is 50% of IP enrichment using rabbit polyclonal CIP29 Antibody. This antibody passes preliminary validation and will be further pursued for primary and secondary validation.
Submitted by
Balaji Sundararaman
Lab
Gene Yeo, UCSD
SARNP (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
IP-WB analysis of K562 whole cell lysate using the SARNP specific antibody, A303-209A. Lanes 1 and 2 are 2.5% of five million whole cell lysate input and 50% of IP enrichment, respectively, using a normal IgG antibody. Lane 3 is 50% of IP enrichment from five million whole cell lysate using the SARNP-specific antibody, A303-209A. The same antibody was used to detect protein levels via Western blot. This antibody passes preliminary validation and will be further pursued for secondary validation. *NOTE* Protein sizes are taken from Genecards.org and are only estimates based on sequence. Actual protein size may differ based on protein characteristics and electrophoresis method used.
Submitter comment
Immunoprecipitation lane (3) exhibits a large deviation from the expected protein size, which we believe is due to performing the characterization using the Jess Western Blotting system. This system has a documented tendency for some proteins to run differently than with more traditional methods due to their compositions and interactions with the electrophoretic components. The image also closely resembles the example image provided by the vendor.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
SARNP (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
IP-Western Blot analysis of HepG2 whole cell lysate using CIP29 specific antibody. Lane 1 is 1% of twenty million whole cell lysate input and lane 2 is 25% of IP enrichment using rabbit normal IgG (lanes under 'IgG'). Lane 3 is 1% of twenty million whole cell lysate input and lane 4 is 10% IP enrichment using rabbit polyclonal anti-CIP29 antibody (lanes under 'CIP29').
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
SARNP (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
IP-Western Blot analysis of K562 whole cell lysate using CIP29 specific antibody. Lane 1 is 1% of twenty million whole cell lysate input and lane 2 is 25% of IP enrichment using rabbit normal IgG (lanes under 'IgG'). Lane 3 is 1% of twenty million whole cell lysate input and lane 4 is 10% IP enrichment using rabbit polyclonal anti-CIP29 antibody (lanes under 'CIP29').
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
SARNP (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot following shRNA against CIP29 in HepG2 whole cell lysate using CIP29 specific antibody. Lane 1 is a ladder, lane 2 is HepG2 non-targeting control knockdown, lane 3 and 4 are two different shRNAs against CIP29. CIP29 protein appears as the green band, Tubulin serves as a control and appears in red.
Submitted by
Xintao Wei
Lab
Brenton Graveley, UConn
SARNP (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot following CRISPR against SARNP in HepG2 whole cell lysate using SARNP specific antibody. Lane 1 is a ladder, lane 2 is HepG2 non-targeting control knockdown, lane 3 and 4 are two different CRISPR against SARNP. SARNP protein appears as the green arrow, Beta-actin serves as a control and appears in red arrow.
Submitted by
Xintao Wei
Lab
Brenton Graveley, UConn
SARNP (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot following CRISPR against SARNP in K562 whole cell lysate using SARNP specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different CRISPR against SARNP. SARNP protein appears as the green arrow, Beta-actin serves as a control and appears in red arrow.
Reviewer comment
Checked with Eric Mendenhall and he agreed with the lab’s explanation: Based on the explanation we got from the lab, these are all CRISPRi based knockouts (unlike the previous RNAi based knockouts) where the lab introduces a stop codon, thus preventing protein synthesis altogether. And as a result, they are able to detect only the protein knockdown rates, but the RNA levels still might not be changing and thus they report XX% k/d using qRT-PCR since they couldn’t even detect anything. This is unlike the RNAi based silencing methods.
Submitted by
Xintao Wei
Lab
Brenton Graveley, UConn